本發(fā)明涉及制藥
技術(shù)領(lǐng)域：
，具體涉及苦參堿肟和一類新的苦參堿衍生物，以及制備方法，以及這些化合物在制備抗腫瘤和殺蟲藥物中的應(yīng)用。
背景技術(shù)：
：苦參是豆科植物的干燥根，含有多種化學(xué)成分。其中，苦參堿是苦參總堿的代表，為羽扇烷寧（白金雀兒堿）的異構(gòu)體，屬于四環(huán)的喹嗪啶類?？鄥A是由豆科植物苦參的干燥根、植株、果實(shí)經(jīng)乙醇等有機(jī)溶劑提取制成的；具有廣泛的藥理作用，如抗腫瘤、抗心律失常、抗炎及抗病毒作用，臨床上主要用于慢性病毒性肝炎的治療，毒副作用小，療效好?？鄥A是一種天然植物性農(nóng)藥,對人畜低毒,是廣譜殺蟲劑，具有觸殺和胃毒作用，對各種作物上的黏蟲、菜青蟲、蚜蟲、紅蜘蛛有明顯的防治效果。肟作為一種重要的有機(jī)反應(yīng)中間體，可以用來制備胺、酰胺、噁唑、吡啶、硝酮、肟酯等化合物，在研發(fā)中常被選為有效的活性基團(tuán)，受到醫(yī)藥和農(nóng)藥行業(yè)的廣泛關(guān)注。肟酯類化合物具有優(yōu)良的殺蟲、殺菌及除草活性，不少品種還具有低毒、低殘留等優(yōu)點(diǎn)。目前該類化合物的分子設(shè)計和合成以及生物活性是研究的熱點(diǎn)，因其具有廣泛的生物活性和多變結(jié)構(gòu)類型而倍受人們的關(guān)注。未見兼具苦參堿的殺蟲活性與抗腫瘤活性和苯甲酰氯類藥物的殺蟲和抗腫瘤活性。技術(shù)實(shí)現(xiàn)要素：本發(fā)明要解決的技術(shù)問題是提供一種用于制備殺蟲和抗腫瘤藥物的苦參堿衍生物及其制備方面。本發(fā)明所述苦參堿衍生物，其結(jié)構(gòu)如下：。本發(fā)明還提供了上述化合物的制備方法，該方法采用如下反應(yīng)路線：。涉及到苦參堿肟LHW-01，具體包括以下技術(shù)方案：（1）LHW-01的制備：具體制備LHW-01的實(shí)驗(yàn)方法：在反應(yīng)瓶中依次加入1.24g（5mmol）苦參堿和20mL甲苯，攪拌下加入5mL雙（三甲基硅基）氨基鈉，于室溫下反應(yīng)10min，然后加入3mL亞硝酸叔丁酯，在50℃下反應(yīng)1.5h（TLC檢測）。冷卻至室溫，真空抽濾，濾液濃縮后經(jīng)硅膠柱層析[洗脫劑：V（石油醚）：V（乙酸乙酯）=1：1]純化得到0.89g白色固體LHW-01。收率65%,m.p.148~153℃；1HNMR(600MHz,CDCl3)δ:15.21(s,1H),4.32(dd,J=12.8,4.4Hz,1H),3.98(dd,J=10.9,7.8,5.3Hz,1H),3.24(t,J=12.8Hz,1H),2.90~2.76(m,2H),2.66(dd,J=15.4,7.4,3.9Hz,1H),2.49(dd,J=15.2,10.9,3.9Hz,1H),2.32~2.20(m,1H),2.07~1.95(m,2H),1.90(dd,J=8.8,6.9Hz,1H),1.81~1.68(m,4H),1.66~1.55(m,3H),1.53~1.41(m,3H),1.33~1.21(m,1H);13CNMR(600MHz,CDCl3)δ:160.94,142.68,63.12,57.08,57.04,53.09,42.48,41.36,35.64,27.57,26.39,26.04,24.76,20.99,20.57.MSm/z:278｛[M+H]+｝；（2）LHW-02的制備：具體制備LHW-02的實(shí)驗(yàn)方法：在反應(yīng)瓶中依次加入1gLHW-01和20mL二氯甲烷，攪拌使其溶解；加入0.5mL吡啶，然后緩慢滴加10%的苯甲酰氯的二氯甲烷溶液，滴畢，升溫回流；TLC檢測，用飽和碳酸氫鈉溶液洗2次，再用水洗2次，至Ph=7，無水硫酸鈉干燥，濃縮，用石油醚和乙酸乙酯混合液進(jìn)行重結(jié)晶，得到LHW-02。淡黃色固體，收率55%,m.p.142~148℃；1HNMR(600MHz,CDCl3)δ:12~8.06(m,2H),7.66~7.61(m,1H),7.50(dd,J=10.8,4.9Hz,2H),4.51(dd,J=12.8,4.5Hz,1H),4.08(ddd,J=10.8,8.0,5.0Hz,1H),3.28(t,J=12.8Hz,1H),3.22(ddd,J=16.3,7.3,4.3Hz,1H),2.92~2.78(m,2H),2.75~2.63(m,1H),2.29(ddt,J=17.2,7.3,4.9Hz,1H),2.01(dtd,J=15.0,12.3,2.8Hz,2H),1.85(s,3H),1.78~1.70(m,3H),1.65(ddd,J=14.3,9.1,4.5Hz,1H),1.61~1.53(m,2H),1.53~1.46(m,3H);13CNMR(600MHz,CDCl3)δ:162.89,158.06,157.85,133.64,129.80,128.65,128.55,63.33,57.12,56.82,52.72,42.89,42.38,35.52,27.56,26.41,23.36,22.42,21.04,20.72;MSm/z:382｛[M+H]+｝；參考化合物L(fēng)HW-02的實(shí)驗(yàn)方法制備化合物L(fēng)HW-03~LHW-24；化合物L(fēng)HW-03~LHW-24結(jié)構(gòu)鑒定數(shù)據(jù)如下：化合物L(fēng)HW-03：白色固體粉末，收率48%，m.p.116~122℃;1HNMR(600MHz,CDCl3)δ7.86(dd,J=7.8,1.1Hz,1H),7.45(td,J=7.6,1.3Hz,1H),7.32–7.24(m,1H),4.49(dd,J=12.8,4.5Hz,1H),4.04(ddd,J=11.0,8.1,5.2Hz,1H),3.26(t,J=12.7Hz,1H),3.14(ddd,J=16.4,7.3,4.3Hz,1H),2.83(dd,J=30.3,11.0Hz,1H),2.67–2.57(m,2H),2.30–2.20(m,1H),1.99(dd,J=22.3,10.0Hz,2H),1.86(d,J=13.8Hz,1H),1.83–1.77(m,1H),1.77–1.65(m,2H),1.65–1.60(m,1H),1.60–1.51(m,1H),1.51–1.41(m,2H)；13CNMR(600MHz,CDCl3)δ:171.07,159.77,148.79,138.0,137.8913.82,130.70,130.44,130.09,128.21,127.27,63.60,56.93,56.80,54.33,52.57,42.71,41.86,42.71,41.86,35.54,27.44,26.34,23.02,21.26,20.81,20.46,19.77;MSm/z:382｛[M+H]+｝；化合物L(fēng)HW-04：黃色固體粉末，收率57%，m.p.170~179℃;1HNMR(600MHz,CDCl3)δ7.83(dd,J=7.7,1.8Hz,1H),7.51(ddd,J=9.0,7.5,1.8Hz,1H),7.03–6.97(m,1H),4.48(dd,J=12.8,4.5Hz,1H),4.08–3.98(m,1H),3.89(s,2H),3.24(dd,J=22.2,9.4Hz,1H),3.19(ddd,J=16.3,7.1,4.3Hz,1H),2.91–2.76(m,1H),2.65–2.54(m,1H),2.24(ddt,J=13.7,7.0,4.9Hz,1H),1.98(dtd,J=15.2,12.3,2.5Hz,1H),1.85(t,J=14.4Hz,1H),1.78(ddd,J=15.5,8.6,3.3Hz,1H),1.75–1.58(m,2H),1.58–1.50(m,1H),1.50–1.41(m,2H);13CNMRδ:163.18,159.14,158.21,157.72,134.13,132.00,120.33,112.08,63.29,57.10,55.99,52.75,42.75,42.39,35.47,27.56,26.36,23.45,22.56,21.04,20.69;MSm/z:412｛[M+H]+｝；化合物L(fēng)HW-05：白色固體粉末，收率41%，m.p.156~165℃;1HNMR(600MHz,CDCl3)δ8.09–8.00(m,1H),7.61–7.54(m,1H),7.27(t,J=5.6Hz,1H),7.16(dt,J=22.7,11.4Hz,1H),4.48(dd,J=12.8,4.5Hz,1H),4.09–4.00(m,1H),3.30–3.16(m,2H),2.83(dd,J=30.8,10.7Hz,2H),2.71–2.60(m,1H),2.30–2.19(m,2H),1.98(dt,J=14.7,7.5Hz,2H),1.87(d,J=13.7Hz,1H),1.80(d,J=9.2Hz,1H),1.77–1.66(m,3H),1.66–1.62(m,1H),1.62–1.51(m,2H),1.51–1.42(m,3H);13CNMR(600MHz,CDCl3)δ:162.52,160.80,158.21,158.00,135.23,135.17,132.78,124.41,124.39,117.09,116.94,63.31,57.12,52.78,42.86,42.37,35.51,27.56,26.39,23.34,22.67,21.04,20.70;MSm/z:400｛[M+H]+｝；化合物L(fēng)HW-06：淡黃色固體粉末，收率52%，m.p.194~203℃;1HNMR(600MHz,DMSO)δ7.76–7.69(m,1H),7.53–7.45(m,1H),7.43–7.35(m,1H),4.21(dd,J=12.6,4.4Hz,1H),3.97–3.84(m,1H),3.10(t,J=12.7Hz,1H),2.85(dd,J=22.5,11.0Hz,1H),2.73(ddd,J=16.5,8.1,4.5Hz,1H),2.40(ddd,J=16.5,9.6,4.7Hz,1H),2.10–1.97(m,2H),1.85(d,J=13.3Hz,1H),1.73–1.64(m,1H),1.62–1.48(m,3H),1.45–1.28(m,2H);13CNMR(600MHz,DMSO)δ:167.65,159.75,149.66,132.42,130.97,130.87,127.57,63.35,56.79,56.74,52.45,42.35,41.35,35.48,27.58,26.20,23.23,20.86,20.42,19.93;MSm/z:416｛[M+H]+｝；化合物L(fēng)HW-07：淡黃色固體粉末，收率45%，m.p.211~215℃;1HNMR(600MHz,DMSO)δ7.69(ddd,J=7.9,3.3,1.4Hz,1H),7.44(td,J=7.5,1.2Hz,1H),7.39(td,J=7.7,1.8Hz,1H),4.21(dd,J=12.6,4.4Hz,1H),3.94–3.84(m,1H),3.10(t,J=12.7Hz,1H),2.84(dd,J=22.4,11.0Hz,1H),2.73(ddd,J=16.5,8.1,4.5Hz,1H),2.51(dt,J=3.5,1.7Hz,1H),2.40(ddd,J=16.5,9.6,4.7Hz,1H),2.10–1.97(m,2H),1.83(t,J=21.9Hz,1H),1.73–1.65(m,1H),1.63–1.48(m,3H),1.45–1.31(m,2H);13CNMR(600MHz,DMSO)δ:168.25,159.74,149.67,134.02,132.42,130.79,128.07,120.21,63.36,56.83,56.77,52.47,42.38,41.38,35.51,27.62,26.24,23.24,20.90,20.45,19.93;MSm/z:460｛[M+H]+｝；化合物L(fēng)HW-08：淡黃色固體粉末，收率35%，m.p.160~168℃;1HNMR(600MHz,CDCl3)δ8.65(s,1H),8.07(dd,J=8.6,1.6Hz,1H),7.98(d,J=8.1Hz,1H),7.91(t,J=8.6Hz,2H),7.66–7.60(m,1H),7.60–7.54(m,1H),4.51(dd,J=12.8,4.5Hz,1H),4.09(ddd,J=11.0,8.0,5.1Hz,1H),3.28(ddd,J=11.6,7.7,4.6Hz,2H),2.90–2.80(m,2H),2.80–2.72(m,1H),2.31(ddt,J=17.3,7.3,4.9Hz,1H),2.01(dtd,J=17.4,12.3,2.5Hz,2H),1.90(d,J=13.8Hz,1H),1.86–1.80(m,1H),1.80–1.69(m,3H),1.67(dd,J=9.2,3.5Hz,1H),1.64–1.53(m,2H),1.53–1.45(m,3H);13CNMR(600MHz,CDCl3)δ:163.11,158.08,157.92,135.77,132.44,131.57,129.40,128.71,128.50,127.85,126.94,125.67,124.99,63.32,62.68,57.13,52.75,42.91,42.36,35.52,27.57,26.43,23.36,22.52,21.06,20.73;MSm/z:432｛[M+H]+｝化合物L(fēng)HW-09：棕色固體粉末，收率32%，m.p.121~128℃;1HNMR(600MHz,CDCl3)δ6.86(d,J=20.9Hz,1H),4.31(d,J=10.7Hz,1H),3.94(d,J=60.3Hz,1H),3.12(d,J=51.0Hz,1H),2.93(ddd,J=21.0,12.5,8.4Hz,1H),2.83(d,J=24.8Hz,1H),2.68–2.52(m,1H),2.43–2.37(m,3H),2.24(d,J=23.7Hz,2H),2.00(s,1H),1.89(d,J=12.9Hz,1H),1.84–1.57(m,4H),1.57–1.35(m,3H);13CNMR(600MHz,CDCl3)δ:167.63,155.62,155.41,139.93,136.38,128.57,128.49,63.30,57.01,53.69,43.33,41.78,35.20,28.32,27.47,26.82,25.95,21.18,20.99,20.58,19.91;MSm/z:424｛[M+H]+｝；化合物L(fēng)HW-10：淡黃色固體粉末，收率42%，m.p.121~136℃;1HNMR(600MHz,CDCl3)δ8.09–7.84(m,2H),7.39(d,J=7.5Hz,1H),7.35(t,J=7.6Hz,1H),4.50(dd,J=12.9,4.5Hz,1H),4.12–4.05(m,1H),3.36-3.25(m,1H),3.02(t,J=14.5Hz,1H),2.96(ddd,J=17.2,8.1,4.7Hz,2H),2.60(ddd,J=17.0,9.6,4.9Hz,1H),2.42(s,3H),2.22(ddd,J=13.1,9.5,4.9Hz,1H),2.06(dd,J=24.9,12.4Hz,2H),1.93(d,J=14.0Hz,1H),1.90-1.78(m,2H),1.77-1.68(m,3H),1.67-1.55(m,2H),1.55-1.45(m,3H),1.28(s,1H);13CNMR(600MHz,CDCl3)δ:171.07,159.77,148.79,138.06,133.82,130.70,130.44,128.21,127.27,63.60,56.93,52.57,42.71,41.86,35.54,27.44,26.34,23.02,21.26,20.81,20.46,19.77;MSm/z:396｛[M+H]+｝；化合物L(fēng)HW-11：白色固體粉末，收率51%，m.p.132~136℃;1HNMR(600MHz,DMSO)δ7.56–7.51(m,1H),7.44(dt,J=8.9,4.5Hz,1H),7.41(t,J=7.9Hz,1H),7.18(ddd,J=8.2,2.7,0.8Hz,1H),4.19(dd,J=12.5,4.3Hz,1H),3.89–3.85(m,1H),3.81(s,3H),3.08(t,J=12.5Hz,1H),2.80–2.69(m,4H),2.41(ddd,J=16.5,9.6,4.7Hz,1H),2.06–2.01(m,1H),1.91–1.83(m,3H),1.65–1.61(m,2H),1.57(dt,J=7.6,3.6Hz,2H),1.56–1.50(m,3H),1.40–1.33(m,3H);13CNMR(600MHz,DMSO)δ:167.84,159.72,159.65,149.68,130.06,122.00,114.37,63.41,57.01,56.94,55.67,52.56,42.52,41.55,40.36,40.22,40.08,39.94,39.81,39.67,39.53,35.67,27.80,26.43,23.21,21.07,20.61,19.92;MSm/z:412｛[M+H]+｝；化合物L(fēng)HW-12：白色固體粉末，收率41%，m.p.126~132℃;1HNMR(600MHz,CDCl3)δ7.89(d,J=7.7Hz,1H),7.77(t,J=12.5Hz,1H),7.43–7.36(m,1H),7.26–7.21(m,1H),4.49(dd,J=13.0,4.5Hz,1H),4.15–4.08(m,1H),3.33(t,J=12.9Hz,1H),3.08(dd,J=38.8,11.0Hz,2H),2.95(ddd,J=17.1,8.0,4.6Hz,1H),2.58(ddd,J=17.0,9.7,4.9Hz,1H),2.21(ddd,J=13.1,9.4,4.8Hz,1H),2.14–2.04(m,2H),1.93(d,J=14.0Hz,1H),1.83(ddd,J=38.5,16.6,8.0Hz,2H),1.78–1.66(m,3H),1.66–1.55(m,2H),1.50(ddd,J=18.2,8.2,3.7Hz,3H);13CNMRδ:169.61,159.75,148.97,129.84,125.75,119.79,116.96,63.62,56.83,52.50,42.59,41.82,35.45,27.31,26.21,23.05,20.65,20.32,19.73;MSm/z:400｛[M+H]+｝；化合物L(fēng)HW-13：白色固體粉末，收率48%，m.p.160~167℃;1HNMR(600MHz,CDCl3)δ8.02(t,J=1.7Hz,1H),7.96(d,J=7.8Hz,1H),7.61–7.56(m,1H),7.43(t,J=7.9Hz,1H),4.48(dd,J=12.8,4.5Hz,1H),4.11–4.00(m,1H),3.26(t,J=12.8Hz,1H),3.23–3.14(m,1H),2.89–2.77(m,2H),2.73–2.63(m,1H),2.28(ddt,J=12.2,7.1,4.9Hz,1H),2.23(s,1H),2.05–1.94(m,2H),1.87(d,J=13.7Hz,1H),1.84–1.78(m,1H),1.78–1.69(m,3H),1.69–1.62(m,1H),1.62–1.51(m,2H),1.51–1.43(m,3H);13CNMR(600MHz,CDCl3)δ:161.71,158.34,157.84,134.80,133.66,130.02,129.68,127.95,63.28,57.11,52.73,42.89,42.35,35.50,27.56,26.40,23.30,22.52,21.04,20.70;MSm/z:416｛[M+H]+｝；化合物L(fēng)HW-14:白色固體粉末，收率35%，m.p.156~165℃;1HNMR(600MHz,CDCl3)δ8.18(t,J=1.7Hz,1H),8.05–7.95(m,1H),7.74(ddd,J=8.0,1.9,1.0Hz,1H),7.37(t,J=7.9Hz,1H),4.48(dd,J=12.8,4.5Hz,1H),4.11–4.00(m,1H),3.25(q,J=12.5Hz,1H),3.22–3.15(m,1H),2.90–2.76(m,2H),2.73–2.61(m,1H),2.33–2.23(m,1H),1.99(dtd,J=14.7,12.3,2.6Hz,2H),1.86(t,J=14.5Hz,1H),1.84–1.78(m,1H),1.78–1.69(m,3H),1.68–1.60(m,1H),1.60–1.52(m,2H),1.52–1.42(m,3H);13CNMR(600MHz,CDCl3)δ:161.59,158.34,157.83,136.59,132.59,130.25,128.40,122.70,63.28,57.12,52.73,42.90,42.36,35.50,27.56,26.41,23.30,22.52,21.04,20.71;MSm/z:460｛[M+H]+｝；化合物L(fēng)HW-15:淡黃色固體粉末，收率45%，m.p.123~130℃;1HNMR(600MHz,DMSO)δ8.23(d,J=7.8Hz,1H),8.18(s,1H),7.99(d,J=7.8Hz,1H),7.76(t,J=7.8Hz,1H),4.23–4.14(m,1H),3.91–3.83(m,1H),3.13–3.02(m,1H),2.83–2.75(m,2H),2.73(ddd,J=16.5,8.2,4.5Hz,1H),2.41(ddd,J=16.5,9.6,4.7Hz,1H),2.06(qd,J=9.3,4.8Hz,1H),2.00–1.89(m,2H),1.85(d,J=13.4Hz,1H),1.70–1.62(m,2H),1.62–1.44(m,4H),1.43–1.28(m,3H);13CNMR(600MHz,DMSO)δ:166.64,159.72,149.70,133.66,130.45,129.88,129.66,129.54,125.96,125.93,125.24,63.42,56.97,56.91,42.47,41.48,35.62,27.75,26.38,23.22,21.03,20.57,19.93;MSm/z:412｛[M+H]+｝；化合物L(fēng)HW-16:16)：白色固體粉末，收率53%，m.p.146~154℃;1HNMR(600MHz,CDCl3)δ:7.96(d,J=8.2Hz,1H),7.27(d,J=8.8Hz,1H),4.49(dd,J=12.8,4.5Hz,1H),4.05(dd,J=10.9,8.0,5.1Hz,1H),3.26(t,J=12.8Hz,1H),3.18(dd,J=16.4,7.3,4.3Hz,1H),2.89~2.77(m,1H),2.71~2.62(m,1H),2.43(s,1H),2.30~2.21(m,1H),1.99(dd,J=14.9,12.3,2.7Hz,1H),1.88(d,J=15.2Hz,2H),1.83~1.77(m,1H),1.77~1.67(m,1H),1.67~1.61(m,1H),1.61~1.51(m,1H),1.51~1.42(m,1H);13CNMR(600MHz,CDCl3)δ:163.98,158.14,157.56,144.53,129.84,129.36,125.70,63.33,57.12,56.82,52.72,42.89,42.38,35.52,27.56,26.41,23.36,22.42,21.77,21.04,20.72;MSm/z:396｛[M+H]+｝；化合物L(fēng)HW-17:白色固體粉末，收率45%，m.p.136~144℃;1HNMR(600MHz,CDCl3)δ:8.19(d,J=8.2Hz,1H),7.75(d,J=8.3Hz,1H),4.48(dd,J=12.8,4.5Hz,1H),4.07(dd,J=10.9,8.1,5.1Hz,1H),3.27(t,J=12.8Hz,1H),3.19(dd,J=16.3,7.2,4.3Hz,1H),2.89~2.78(m,1H),2.69(dd,J=16.0,11.0,4.7Hz,1H),2.29(dd,J=17.2,7.2,4.9Hz,1H),2.02~1.98(m,1H),1.95(s,1H),1.87(d,J=13.6Hz,1H),1.84~1.78(m,1H),1.77~1.69(m,2H),1.68~1.61(m,1H),1.60~1.52(m,1H),1.48(d,J=6.9,4.1Hz,2H);13CNMR(600MHz,CDCl3)δ:161.78,158.48,157.78,131.84,130.18,125.72,125.69,63.72,57.12,57.11,52.73,42.92,42.39,35.51,27.55,26.41,23.31,22.52,21.04,20.71;MSm/z:450｛[M+H]+｝；化合物L(fēng)HW-18：淡黃色固體粉末，收率49%，m.p.156~162℃;1HNMR(600MHz,CDCl3)δ:8.14~8.07(m,1H),7.21~7.13(m,1H),4.50(dd,J=12.8,4.5Hz,1H),4.12~4.04(m,1H),3.28(t,J=12.8Hz,1H),3.19(dd,J=16.4,7.3,4.3Hz,1H),2.85(dd,J=31.5,10.9Hz,1H),2.73~2.62(m,1H),2.34~2.24(m,1H),2.02(d,J=16.2,12.0Hz,1H),1.87(s,1H),1.83(d,J=11.9Hz,1H),1.74(dd,J=14.1,10.2,4.3Hz,2H),1.64(dd,J=24.4,10.1Hz,1H),1.61~1.53(m,1H),1.53~1.45(m,2H);13CNMR(600MHz,CDCl3)δ:161.99,157.98,157.88,132.44,132.39,116.01,115.87,63.30,57.12,56.89,52.71,42.92,42.38,35.52,27.55,26.41,23.31,22.42,21.04,20.71;MSm/z:400｛[M+H]+｝；化合物L(fēng)HW-19:白色固體粉末，收率49%，m.p.140~155℃;1HNMR(600MHz,CDCl3)δ:8.04~7.98(m,1H),7.49~7.43(m,1H),4.49(dd,J=12.8,4.5Hz,1H),4.07(s,1H),3.28(t,J=12.7Hz,1H),3.18(dd,J=16.4,7.3,4.3Hz,1H),2.85(dd,J=31.8,10.4Hz,1H),2.74~2.65(m,1H),2.33~2.24(m,1H),2.04(s,1H),1.88(d,J=13.7Hz,1H),1.82(d,J=9.5Hz,1H),1.78~1.70(m,2H),1.69~1.62(m,1H),1.57(dd,J=18.4,9.5,4.2Hz,1H),1.53~1.45(m,2H)；13CNMR(600MHz,CDCl3)δ:162.15,157.97,157.94,140.24,131.14,129.06,126.90,63.28,57.10,56.88,52.72,42.92,42.35,35.51,27.54,26.40,23.27,22.42,21.02,20.69;MSm/|z:41｛[M+H]+｝；化合物L(fēng)HW-20:淡黃色固體粉末，收率40%，m.p.136~141℃;1HNMR(600MHz,CDCl3)δ:7.93(d,J=8.5Hz,1H),7.93(d,J=8.5Hz,1H),7.63(d,J=8.5Hz,1H),7.63(d,J=8.5Hz,1H),4.49(d,J=9.5Hz,1H),4.06(s,1H),3.27(t,J=12.4Hz,1H),3.21~3.11(m,1H),2.91~2.74(m,1H),2.73~2.56(m,1H),2.25(t,J=16.2Hz,1H),2.16(s,1H),2.01(t,J=18.4Hz,1H),1.87(d,J=13.1Hz,1H),1.80(s,1H),1.72(s,3H),1.64(d,J=10.4Hz,1H),1.57(d,J=8.9Hz,1H),1.48(s,2H);13CNMR(600MHz,CDCl3)δ:162.21,158.06,157.92,132.06,131.69,131.23,128.91,127.38,63.31,57.08,56.77,52.69,42.87,42.33,35.46,27.51,26.36,23.31,22.44,20.99,20.66;MSm/z:461｛[M+H]+｝；化合物L(fēng)HW-21:淡黃色固體粉末，收率55%，m.p.158~170℃;1HNMR(600MHz,CDCl3)δ:8.03(t,J=5.7Hz,1H),6.96(t,J=5.7Hz,1H),4.49(dd,J=12.8,4.5Hz,1H),4.05(dd,J=11.0,8.0,5.1Hz,1H),3.88(s,2H),3.25(t,J=12.7Hz,1H),3.18(dd,J=16.3,7.3,4.3Hz,1H),2.83(dd,J=31.6,10.9Hz,1H),2.71~2.62(m,1H),2.31~2.20(m,1H),2.05~1.93(m,1H),1.93~1.83(m,2H),1.80(dd,J=17.3,8.6Hz,1H),1.77~1.68(m,2H),1.68~1.59(m,1H),1.55(dd,J=19.0,9.9,4.4Hz,1H),1.52~1.40(m,2H);13CNMR(600MHz,CDCl3)δ:163.91,162.63,158.17,157.41,131.93,120.70,113.95,63.34,57.14,57.01,22.53,52.71,42.88,42.37,35.53,27.57,26.43,23.36,22.36,21.06,20.74;MSm/z:412｛[M+H]+｝；化合物L(fēng)HW-22:白色固體粉末，收率56%，m.p.122~127℃;1HNMR(600MHz,CDCl3)δ8.02–7.98(m,1H),7.52–7.47(m,1H),4.48(dd,J=12.8,4.5Hz,1H),4.10–4.01(m,1H),3.31–3.22(m,1H),3.21–3.13(m,1H),2.90–2.78(m,1H),2.66(tdd,J=11.0,9.1,4.2Hz,1H),2.29–2.23(m,1H),2.16(s,1H),1.99(dtd,J=14.6,12.2,2.6Hz,1H),1.87(d,J=13.8Hz,1H),1.80(ddd,J=16.5,9.1,2.7Hz,1H),1.76–1.67(m,1H),1.63(ddd,J=21.7,7.0,3.5Hz,1H),1.60–1.51(m,1H),1.51–1.43(m,1H),1.36–1.33(m,1H);13CNMR(600MHz,CDCl3)δ:163.03,158.21,157.54,157.41,129.71,125.66,125.56,63.31,57.11,52.73,42.92,42.35,35.53,35.21,31.07,27.55,26.40,23.27,22.31,21.03,20.70;MSm/z:438｛[M+H]+｝；化合物L(fēng)HW-23:23)：淡黃色固體粉末，收率56%，m.p.120~135℃;1HNMR(600MHz,CDCl3)δ:8.17~8.11(m,2H),7.72~7.68(m,2H),7.66~7.59(m,2H),7.51~7.46(m,2H),7.44~7.38(m,1H),4.50(dd,J=12.8,4.5Hz,1H),4.10~4.03(m,1H),3.27(t,J=12.8Hz,1H),3.22(dd,J=16.4,7.3,4.3Hz,1H),2.84(dd,J=32.1,10.9Hz,2H),2.75~2.64(m,1H),2.32~2.24(m,1H),2.00(dd,J=14.9,12.2,2.6Hz,2H),1.88(d,J=13.6Hz,1H),1.81(dd,J=13.5,10.0Hz,1H),1.64(dd,J=15.4,11.8Hz,1H),1.61~1.52(m,2H),1.48(d,J=13.0,4.5Hz,3H);13CNMR(600MHz,CDCl3)δ:162.77,158.07,157.91,142.42,139.80,130.34,129.80,129.01,128.38,127.32,63.35,57.13,56.95,52.72,42.88,42.62,42.40,35.52,27.56,26.42,23.41,22.47,21.05,20.72;MSm/z:458｛[M+H]+｝；化合物L(fēng)HW-24:淡黃色固體粉末，收率29%，m.p.152~156℃;1HNMR(600MHz,DMSO)δ8.90–8.84(m,1H),8.13(dd,J=9.6,3.5Hz,1H),8.00(d,J=7.9Hz,1H),7.66–7.60(m,1H),7.57(td,J=7.0,3.1Hz,1H),4.19(dd,J=12.5,4.0Hz,1H),3.89–3.82(m,1H),3.07(t,J=12.6Hz,1H),2.81–2.73(m,1H),2.72(ddd,J=16.7,8.2,4.7Hz,1H),2.41(tdd,J=14.5,10.0,4.5Hz,1H),2.03(ddd,J=34.5,19.4,15.2Hz,1H),1.96–1.86(m,1H),1.81(d,J=11.2Hz,1H),1.67–1.46(m,2H),1.40–1.28(m,1H);13CNMR(600MHz,DMSO)δ:169.48,160.73,159.74,149.66,133.94,132.97,131.19,130.02,129.00,127.81,126.54,126.14,125.35,63.34,56.91,52.51,42.46,41.48,40.38,39.54,35.60,27.72,26.33,23.21,21.00,20.54,19.93.MSm/z:432｛[M+H]+｝；通過本發(fā)明制備的上述苦參堿肟酯類衍生物，其構(gòu)成純度在99%以上。以下表1給出了制備目標(biāo)化合物的結(jié)構(gòu)式：本發(fā)明還包括，所述苦參堿衍生物在制備抗腫瘤藥物中的應(yīng)用。優(yōu)選的，所述腫瘤為肝癌、肺癌、胃癌細(xì)和乳腺細(xì)。以下通過實(shí)驗(yàn)進(jìn)一步說明本發(fā)明的有益效果。藥理實(shí)驗(yàn)：人類肝癌細(xì)胞(Bel-7402)和人類乳腺癌細(xì)胞(MCF-7)培養(yǎng)在％胎牛血清和1％青霉素-鏈霉素的RPMI1640培養(yǎng)基中；人類胃癌細(xì)胞(SGC-7901)培養(yǎng)在含10％FBS和1％青霉素的DMEM高糖培養(yǎng)基中；人類肺癌細(xì)胞(H460)培養(yǎng)在含10％FBS和1％青霉素-鏈霉素的McCoy5A培養(yǎng)基中；所有細(xì)胞都置于37℃，5％CO2的細(xì)胞培養(yǎng)箱中。用MTT法進(jìn)行測試，將苦參堿、苦參堿肟酯類化合物溶解后用基礎(chǔ)培養(yǎng)基稀釋成母液。取0.5x106個對數(shù)生長期的以上細(xì)胞株，接種于96孔板中，每孔100μL（5000個細(xì)胞），設(shè)空白組、對照組以及給藥組（50μM）,每組設(shè)4個平行孔。置于37℃、5%（V/V）CO2的培養(yǎng)箱中，24h后加入同一濃度的藥物，繼續(xù)培養(yǎng)72h，終止前除去上清，每空加入30μL5mg/mLMTT，培養(yǎng)箱孵育4h后除去MTT并加入100μLDMSO/孔，置搖床上低速振蕩10min，使結(jié)晶物充分溶解，運(yùn)用酶標(biāo)儀在490nm波長處測定各孔吸光度（OD）值，計算細(xì)胞增殖抑制率，細(xì)胞增殖抑制率=(1-樣品OD值/對照組OD值)×100%。上述實(shí)驗(yàn)重復(fù)3次，并用Blies法計算出IC50值，組間進(jìn)行比較采用的是t檢驗(yàn)法，P<0.05為差異有顯著性。實(shí)驗(yàn)結(jié)果：表2目標(biāo)化合物對Bel-7402，SGC-7901，MCF-7和H460細(xì)胞增殖的抑制藥理實(shí)驗(yàn)結(jié)果表明：合成的系列苦參堿衍生物均有抗腫瘤活性作用，在以苦參堿為對照藥，與合成的LHW系列化合物同時給藥的情況下，通過對比它們的抑制率發(fā)現(xiàn)，所有的化合物的活性均優(yōu)于對照藥苦參堿，LHW-03、LHW-08、LHW-11、LHW-13、LHW-16、LHW-17、LHW-18、LHW-19、LHW-22、LHW-23均小于50μM，其中LHW-03、LHW-17、LHW-22均小于10μM，活性有了明顯的提高。本發(fā)明在于提供了一種有望成為高效低毒的抗腫瘤藥物，給廣大癌癥患者多一種治療的選擇。本發(fā)明還包括，所述苦參堿衍生物在殺蟲藥物中的應(yīng)用。優(yōu)選的，所述害蟲為紅蜘（Panonychuscitri），健康、活潑、大小一致的3齡幼蟲，由廣西田園生化股份有限公司生測室提供。蚜蟲（Aphidoidea），健康、活潑、大小一致的若蟲，由廣西田園生化股份有限公司生測室提供。以下通過實(shí)驗(yàn)進(jìn)一步說明本發(fā)明的有益效果。藥理實(shí)驗(yàn)：1、紅蜘蛛觸殺活性試驗(yàn)：采用葉片浸漬法，將所合成的苦參堿肟酯類化合物以二甲基亞砜（DMSO）和10%吐溫80（93.5%）配成母液，用水稀釋到200ppm的藥液，將無污染的新鮮酢漿草逐一在配好的稀釋液中浸漬5s，拿出置吸水紙上晾干，然后接入試蟲，任其取食帶藥葉片，以含有二甲基亞砜的10%吐溫80（93.5%）水溶液作為空白對照，盒蓋后置于室溫的環(huán)境下，48h檢查結(jié)果。死亡標(biāo)準(zhǔn)：以毛筆尖觸其蟲體,無任何反應(yīng)者為死亡。死亡率和有效率分別用公式1-1和公式1-2計算。校正死亡率和校正有效率使用Abbott公式進(jìn)行校正。公式1-1：公式1-2：2、蚜蟲觸殺活性試驗(yàn)：同樣采用葉片浸漬法，將所合成的苦參堿肟酯類化合物以二甲基亞砜（DMSO）和10%吐溫80（93.5%）配成母液，用水稀釋到200ppm的藥液，將無污染的新鮮小豆苗逐一在配好的稀釋液中浸漬5s，拿出置吸水紙上晾干，然后接入試蟲，任其取食帶藥葉片，以含有二甲基亞砜的10%吐溫80（93.5%）水溶液作為空白對照，盒蓋后置于室溫的環(huán)境下，48h檢查結(jié)果。死亡標(biāo)準(zhǔn)：以毛筆尖觸其蟲體,無任何反應(yīng)者為死亡。死亡率和有效率分別用公式1-1和公式1-2計算。校正死亡率和校正有效率使用Abbott公式進(jìn)行校正。實(shí)驗(yàn)結(jié)果：本實(shí)驗(yàn)選取了兩種田間常見害蟲—紅蜘蛛和蚜蟲為實(shí)驗(yàn)對象，采用葉片浸漬法測試苦參堿肟酯類衍生物對害蟲的觸殺活性（死亡率），結(jié)果如表3所示。表3.目標(biāo)化合物對害蟲的觸殺活性編號對紅蜘蛛觸殺活性對蚜蟲觸殺活性編號對紅蜘蛛觸殺活性對蚜蟲觸殺活性LHW-01--LHW-130.360.30LHW-020.550.61LHW-140.530.61LHW-030.670.96LHW-150.850.91LHW-040.100.93LHW-160.930.80LHW-050.100.95LHW-170.970.99LHW-060.990.94LHW-180.960.94LHW-070.880.93LHW-190.960.84LHW-080.970.74LHW-200.140.52LHW-090.930.95LHW-210.980.13LHW-100.840.81LHW-220.000.12LHW-110.970.22LHW-230.960.08LHW-120.080.70LHW-240.000.82阿維菌素1.000.99苦參堿0.210.48藥理實(shí)驗(yàn)結(jié)果表明：合成的系列苦參堿衍生物均有抗腫瘤活性作用，在以苦參堿和阿維菌素為對照藥，與合成的LHW系列化合物同時給藥的情況下，通過對比它們的殺蟲死亡率發(fā)現(xiàn)，在給藥濃度為200ppm時，較苦參堿而言，對紅蜘蛛具有較好的觸殺活性。其中有10個目標(biāo)化合物活性達(dá)到了90%以上，9個化合物與對照組阿維菌素的觸殺活性接近，LHW-06的活性達(dá)到了99%。在目標(biāo)化合物對蚜蟲的觸殺活性測試中，其中有9個目標(biāo)化合物活性達(dá)到了90%以上，化合物與對照組阿維菌素的觸殺活性接近，其中LHW-17的活性達(dá)到了99%。本發(fā)明在于提供了一種有望成為高效低毒無污染的殺蟲藥物，給廣大農(nóng)民多一種治療的選擇。當(dāng)前第1頁1 2 3